The introduction of consensus -35 (TTGACA) and -10 (TATAAT) hexamers and a TG motif into the Lactobacillus acidophilus ATCC 4356 wild-type slpA promoter resulted in significant improvements (4.3-, 4.1-, and 10.7-fold, respectively) in transcriptional activity in Lactobacillus fermentum BR11. In contrast, the same changes resulted in decreased transcription in Lactobacillus rhamnosus GG. The TG motif was shown to be important in the context of weak -35 and -10 hexamers (L. fermentum BR11) or a consensus - 10 hexamer (L. rhamnosus GG). Thus, both strain- and context-dependent effects are critical factors influencing transcription in Lactobacillus.